The oncoprotein gankyrin binds to MDM2/HDM2, enhancing ubiquitylation and degradation of p53.
Gankyrin is an ankyrin repeat oncoprotein commonly overexpressed in hepatocellular carcinomas. Gankyrin interacts with the S6 proteasomal ATPase and accelerates the degradation of the tumor suppressor Rb. We show here that gankyrin has an antiapoptotic activity in cells exposed to DNA damaging agents. Downregulation of gankyrin induces apoptosis in cells with wild-type p53. In vitro and in vivo experiments revealed that gankyrin binds to Mdm2, facilitating p53-Mdm2 binding, and increases ubiquitylation and degradation of p53. Gankyrin also enhances Mdm2 autoubiquitylation in the absence of p53. Downregulation of gankyrin reduced amounts of Mdm2 and p53 associated with the 26S proteasome. Thus, gankyrin is a cofactor that increases the activities of Mdm2 on p53 and probably targets polyubiquitylated p53 into the 26S proteasome.


Bioresponsive drug delivery systems in intestinal inflammation: State-of-the-art and future perspectives.
Oral colon-specific delivery systems emerged as the main therapeutic cargos by making a significant impact in the field of modern medicine for local drug delivery in intestinal inflammation. The site-specific delivery of therapeutics (aminosalicylates, glucocorticoids, biologics) to the ulcerative mucus tissue can provide prominent advantages in mucosal healing (MH). Attaining gut mucosal healing and anti-fibrosis are main treatment outcomes in inflammatory bowel disease (IBD). The pharmaceutical strategies that are commonly used to achieve a colon-specific drug delivery system include time, pH-dependent polymer coating, prodrug, colonic microbiota-activated delivery systems and a combination of these approaches. Amongst the different approaches reported, the use of biodegradable polysaccharide coated systems holds great promise in delivering drugs to the ulcerative regions. The present review focuses on major physiological gastro-intestinal tract challenges involved in altering the pharmacokinetics of delivery systems, pathophysiology of MH and fibrosis, reported drug-polysaccharide cargos and focusing on conventional to advanced disease responsive delivery strategies, highlighting their limitations and future perspectives in intestinal inflammation therapy.


Reproducibility of the interpretation of hysterosalpingography in the diagnosis of tubal pathology.
The aim of the study was to estimate the inter- and intra-observer reproducibility of the interpretation of hysterosalpingography (HSG) in the diagnosis of tubal pathology, and associate reproducibility with diagnostic accuracy. Four observers evaluated 143 HSGs twice, on proximal tubal obstruction, distal tubal obstruction, hydrosalpinx and peritubal adhesions. Diagnostic laparoscopy with chromopertubation was considered to be the reference strategy. Reproducibility (inter- and intra-observer agreement) was expressed in terms of kappa-values. Accuracy was expressed in terms of sensitivity, specificity and likelihood ratios. kappa-values for reproducibility between observers were almost perfect for proximal obstruction, substantial for distal obstruction and hydrosalpinx, and moderate for adhesions. kappa-values for reproducibility within observers were almost perfect for proximal obstruction and substantial for distal obstruction, hydrosalpinx and adhesions. HSG had a high specificity for proximal obstruction, but a low sensitivity. Distal obstruction, absence of hydrosalpinx and adhesions had a poor accuracy. The likelihood ratio for the presence of hydrosalpinx was high. In conclusion, proximal tubal obstruction detected on HSG changes the pre-test probability of proximal tubal obstruction from 16 to 50%. Proximal tubal patency detected on HSG changes the pre-test probability of proximal tubal patency from 16 to 9%. It is unlikely that a lack of reproducibility of the interpretation of proximal tubal patency is responsible for the low sensitivity; alternative explanations are artefacts occurring while performing HSG or an imperfect reference strategy diagnostic laparoscopy. HSG is of limited use in diagnosing distal tubal obstruction and hydrosalpinx, and has no value in the detection of peritubal adhesions.


Making Waves: Principles for the Design of Sustainable Household Water Treatment.
Household water treatment (HWT) represents a key engineering intervention for billions of people's daily and emergency water needs. However, established HWT practices experience different challenges to achieve sustainability. Here, five essential principles are identified and discussed for the design of sustainable HWT, including treatment capability, environmental friendliness, user experience, economic viability, and social acceptance. A well-implemented HWT, with a consideration of energy resilience, relies heavily on advanced materials, innovative technologies, and/or creative designs capable of reliably abating both traditional and emerging contaminants, while minimizing undesirable chemical leaching and the formation of harmful disinfection byproducts. Environmental friendliness can be pursued by appreciably reducing environmental and energy footprints and properly managing HWT residuals. Meanwhile, a user-centered design approach and economic consideration need to be integrated into the HWT development to improve end users' willingness to use and pay, respectively. Finally, social acceptance should be intertwined by fostering public acceptance and market adoption for strengthening commercial viability of new-generation HWT products. Given that these principles span across different domains, multidisciplinary collaboration is required throughout different stages of the HWT technology innovation.


Effects of caffeine consumption in patients with chronic hepatitis C: A systematic review and meta-analysis.
Increased caffeine consumption has been associated with a decreased risk of liver enzyme elevation, cirrhosis, and hepatocellular carcinoma. However, few studies have assessed these effects in patients with chronic hepatitis C; therefore, we conducted a systematic review and meta-analysis to investigate the impact of caffeine consumption in patients with chronic hepatitis C infection. We performed a comprehensive search of the databases of the Cochrane Central Register of Controlled Trials, MEDLINE, and EMBASE from inception through November 2015. The inclusion criterion was observational studies' assessment of the impact of caffeine consumption in adult patients with chronic hepatitis C. Eleven studies were included for full article review, and data was extracted from five observational studies for meta-analysis. The pooled odds ratio of advanced hepatic fibrosis in patients who had higher caffeine intake was 0.39 (95% confidence interval 0.21-0.72, P=0.003) compared with lower caffeine intake group. The statistical between-study heterogeneity was moderate with an I2 of 70%. Our meta-analysis demonstrated that caffeine intake is significantly associated with decreased odds of advanced hepatic fibrosis in patients with chronic hepatitis C. Future prospective studies assessing the optimal dose and preparation of caffeinated beverages for prevention of hepatic fibrosis are needed.


A comparison of the palliative care needs of patients dying from chronic respiratory diseases and lung cancer.
This paper reports on data from the Regional Study of Care for the Dying, conducted in 1990, and compares symptoms, care and service utilization for patients with chronic lung diseases (CLD) and lung cancer (LC) in the final 12 months of life. Post-bereavement structured interviews were conducted with informal carers of 449 LC patients and 87 CLD patients. The LC patients were significantly younger than those with CLD (P = 0.001) and these respondents were more likely to have been a spouse (P = 0.034). No differences were found in the mean number of symptoms reported by the two groups in the final year or week of life, although the CLD patients were more likely to have experienced these symptoms for longer. Significantly more patients with CLD than LC experienced breathlessness in the final year (94% CLD vs 78% LC, P < 0.001) and final week (91% CLD vs 69% LC, P < 0.001) of life. Significantly more LC patients were reported to have experienced anorexia (76% LC vs 67% CLD, P = 0.06) and constipation (59% LC vs 44% CLD, p = 0.01) in the final year of life. There were no differences in general practitioner use, but LC patients were reported to have received more help from district nurses (52% LC vs 39% CLD, P = 0.025) and from a palliative care nurse (29% LC vs 0% CLD, P < 0.001). More CLD patients were reported to have received help from social services (29% CLD vs 18% LC, P = 0.037). LC patients were reported to be more likely to have known they might die (76% LC vs 62% CLD, P = 0.003) and to have been told this by a hospital doctor (30% LC vs 8% CLD, P = 0.001). Among those that knew, LC patients were told earlier prior to death than CLD patients. This study suggests that patients with CLD at the end of life have physical and psychosocial needs at least as severe as patients with lung cancer.


Mitochondrial Dysfunction Prevents Repolarization of Inflammatory Macrophages.
Macrophages are innate immune cells that adopt diverse activation states in response to their microenvironment. Editing macrophage activation to dampen inflammatory diseases by promoting the repolarization of inflammatory (M1) macrophages to anti-inflammatory (M2) macrophages is of high interest. Here, we find that mouse and human M1 macrophages fail to convert into M2 cells upon IL-4 exposure in vitro and in vivo. In sharp contrast, M2 macrophages are more plastic and readily repolarized into an inflammatory M1 state. We identify M1-associated inhibition of mitochondrial oxidative phosphorylation as the factor responsible for preventing M1→M2 repolarization. Inhibiting nitric oxide production, a key effector molecule in M1 cells, dampens the decline in mitochondrial function to improve metabolic and phenotypic reprogramming to M2 macrophages. Thus, inflammatory macrophage activation blunts oxidative phosphorylation, thereby preventing repolarization. Therapeutically restoring mitochondrial function might be useful to improve the reprogramming of inflammatory macrophages into anti-inflammatory cells to control disease.


Genetic polymorphism of the human cytochrome CYP2A13 in a French population: implication in lung cancer susceptibility.
The human cytochrome CYP2A13, which is mainly expressed in the respiratory tract, has been shown to be highly efficient in vitro in the metabolism of tobacco-smoke carcinogens and procarcinogens such as 4-methylnitroso-1-(3-pyridyl)-1-butanone (NNK). In order to investigate the extent of CYP2A13 genetic polymorphism in a French Caucasian population of 102 individuals, a screening for sequence variations in the 5'-untranslated and protein encoding regions of its gene was performed using a polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) strategy. Six polymorphisms in the coding region were identified, including two rare missense mutations (C474G or Asp158Glu, G967T or Val323Leu) and one nonsense mutation (Arg101Stop). This deleterious mutation, the most frequent (5%) in our population, presumably encodes a severely truncated protein. The influence of the nonsense mutation in lung cancer susceptibility was examined by PCR-SSCP using peripheral blood DNA from 204 cases of lung cancer and 201 controls. The CYP2A13*7 allele, which harbours the C301T mutation, was present in 2.0% of controls and 3.4% of cases. However, multivariate analysis showed an elevated risk for small cell lung cancer in subjects heterozygous for the null allele (odds ratio OR=9.9; 95% confidence interval CI=1.9-52.2). This increased risk was not linked to other histological types of lung cancer.


Comparison of Anaplasma and Ehrlichia species-specific peptide ELISAs with whole organism-based immunofluorescent assays for serologic diagnosis of anaplasmosis and ehrlichiosis in dogs.
To compare the performance of 5 synthetic peptide-based ELISAs with that of 3 commercially available immunofluorescent assays (IFAs) for serologic diagnosis of anaplasmosis and ehrlichiosis in dogs. A convenience set of 109 serum samples obtained before and at various times after inoculation for 23 dogs that were experimentally infected with Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Ehrlichia chaffeensis, or Ehrlichia ewingii and 1 uninfected control dog in previous studies. All serum samples were assessed with 5 synthetic peptide-based ELISAs designed to detect antibodies against A phagocytophilum, A platys, E canis, E chaffeensis, and E ewingii and 3 whole organism-based IFAs designed to detect antibodies against A phagocytophilum, E canis, and E chaffeensis. The species-specific seroreactivity, cross-reactivity with the other tick-borne pathogens (TBPs), and diagnostic sensitivity and specificity were calculated for each assay and compared among assays. All serum samples obtained from dogs experimentally infected with a TBP yielded positive results on a serologic assay specific for that pathogen. In general, sensitivity was comparable between ELISAs and IFAs and tended to increase with duration after inoculation. Compared with the IFAs, the corresponding ELISAs were highly specific and rarely cross-reacted with antibodies against other TBPs. Results suggested that peptide-based ELISAs had enhanced specificity relative to whole organism-based IFAs for detection of antibodies against Anaplasma and Ehrlichia spp, which should facilitate accurate diagnosis and may help detect dogs coinfected with multiple TBPs.


Specific findings in distortion product otoacoustic emissions and growth functions with chronic tinnitus.
Chronic tinnitus has a very high prevalence in industrialized countries. Latest studies found chronic tinnitus in 4% of the German population, with almost 2% suffering severely in their daily life. Because no curative therapeutic approach is available--neither pharmacological nor surgical--the main focus lies in treatments that enhance the habituation of tinnitus. Habituation occurs in almost 50% of affected patients as a normal process, leading to a complete compensation. This finding is based on the ability of the auditory perception to habituate random noise and focus on important acoustic information. According to our audiological data, 90% of tinnitus patients have deficits in inner-ear function as a generator of tinnitus, mainly in the outer hair cells. This occurrence can be verified by registration of distortion products of otoacoustic emissions. Thus, the main origin of tinnitus is peripheral, and most patients suffer from accompanying hearing loss, even though it is sometimes mild or subjectively not even noticed. In almost 50% of our patients, we find hyperfunction of outer hair cells, again recorded via distortion products of otoacoustic emissions and their growth functions. Normal efferent reduction of distortion products through contralateral acoustic stimulation does not take place in most tinnitus patients. This finding shows that central auditory functions are also disturbed in chronic tinnitus patients, leading to reduced efferent effects on the hair cells and thus impeding habituation. Tests to verify these more central pathological findings have yet to be developed. We have data on diminished ability to distinguish stimuli from random noise by bilateral sound processing: The so-called bilateral masking difference test results are pathological in almost 30% of patients suffering from chronic tinnitus. We concluded from our audiological data that chronic tinnitus is primarily a cochlear dysfunction, but habituation is impeded by accompanying or consecutive deficits of the central auditory pathway. Regarding therapeutic approaches, these central functions can be trained by hearing therapy, as we know from patients' rehabilitation.


Fetal development of the segment-specific papillary body in the equine hoof.
Fetal development of the unique papillary body and its localized peculiarities in the equine hoof are described based on the study of 51 fetuses, nine newborn foals, and five adult horses. The shape and dimensions of the dermal papillae and lamellae have a formative influence on the structure and physical quality of the corneous hoof capsule with its horn tubules and lamellae. The size and arrangement of these horn structures determine the mechanical quality of hoof horn. Proper horn quality is a prerequisite for the various functions of the hoof capsule, such as protecting the living dermis supporting the hoof capsule, shock absorption, and formation of the suspensory apparatus of the distal phalanx. Development of the segment-specific papillary body is initiated by the increasing mitotic activity of the epidermal cells invaginating the dermal surface, thus forming dermal microridges. These microridges are transformed into single dermal papillae, which are arranged in rows, or enlarged to become primary and secondary dermal lamellae. The formation of a segment-specific papillary body enables the increasing keratinization ratio in the hoof epidermis and the formation of the characteristic tubular and lamellar horn responsible for the special mechanical properties of hoof horn.


A psychometric typology of U.K. domestic violence offenders.
A number of studies have described subtypes of domestically violent men, and the heterogeneity of domestically violent men is well established. The aim of the current study was to investigate the distribution of subtypes using psychometric measures in convicted domestically violent offenders in England. Four subtypes of offenders were identified: low pathology, borderline, narcissistic, and antisocial. These subtypes were broadly comparable with the family-only, dysphoric/borderline, and generally violent/antisocial types proposed by Holtzworth-Munroe and Stuart. The majority of the sample (60%) best fit the generally violent/antisocial profile. However, the reliance on psychometric measurement and lack of corroborative evidence from partners means that further research is necessary to test these findings.


[Registration of ethnicity allowed with conditions].
Registration of an individual's ethnicity is, in the light of the potential risks of stigmatization and discrimination, rightfully considered a sensitive issue. Traditionally, privacy legislation offers special legal protection in the collection, registration etc. of data relating to race and ethnic background. However, if it can be established that registration of ethnicity is necessary for providing good care, registration is lawful. However, registration for health research purposes requires the explicit consent of the persons involved.


Response of human REV1 to different DNA damage: preferential dCMP insertion opposite the lesion.
REV1 functions in the DNA polymerase zeta mutagenesis pathway. To help understand the role of REV1 in lesion bypass, we have examined activities of purified human REV1 opposite various template bases and several different DNA lesions. Lacking a 3'-->5' proofreading exonuclease activity, purified human REV1 exhibited a DNA polymerase activity on a repeating template G sequence, but catalyzed nucleotide insertion with 6-fold lower efficiency opposite a template A and 19-27-fold lower efficiency opposite a template T or C. Furthermore, dCMP insertion was greatly preferred regardless of the specific template base. Human REV1 inserted a dCMP efficiently opposite a template 8-oxoguanine, (+)-trans-anti-benzo[a]pyrene-N2-dG, (-)-trans-anti-benzo[a]pyrene-N2-dG and 1,N6-ethenoadenine adducts, very inefficiently opposite an acetylaminofluorene-adducted guanine, but was unresponsive to a template TT dimer or TT (6-4) photoproduct. Surprisingly, the REV1 specificity of nucleotide insertion was very similar in response to different DNA lesions with greatly preferred C insertion and least frequent A insertion. By combining the dCMP insertion activity of human REV1 with the extension synthesis activity of human polymerase kappa, bypass of the trans-anti-benzo[a]pyrene-N2-dG adducts and the 1,N6-ethenoadenine lesion was achieved by the two-polymerase two-step mechanism. These results suggest that human REV1 is a specialized DNA polymerase that may contribute to dCMP insertion opposite many types of DNA damage during lesion bypass.


Spot-scan imaging in transmission electron microscopy.
The determination of the structure of proteins and other organic materials by transmission electron microscopy is a rapidly developing field. Obtaining high-resolution images of these radiation-sensitive specimens has, until recently, been problematic. The development of spot-scan imaging, in which the electron beam is focused to a spot with a diameter of about 1000 angstroms and moved over the specimen to record the image, has overcome some of the most severe problems, which result from beam-induced motion of the specimen and its image. Elimination of this motion greatly enhances the contrast of high-resolution features of the image and promises a significant increase in the speed with which future structural work can be accomplished.


Atomic layer deposited second-order nonlinear optical metamaterial for back-end integration with CMOS-compatible nanophotonic circuitry.
We report the fabrication of artificial unidimensional crystals exhibiting an effective bulk second-order nonlinearity. The crystals are created by cycling atomic layer deposition of three dielectric materials such that the resulting metamaterial is noncentrosymmetric in the direction of the deposition. Characterization of the structures by second-harmonic generation Maker-fringe measurements shows that the main component of their nonlinear susceptibility tensor is about 5 pm/V, which is comparable to well-established materials and more than an order of magnitude greater than reported for a similar crystal [Appl. Phys. Lett.107, 121903 (2015)APPLAB0003-695110.1063/1.4931492]. Our demonstration opens new possibilities for second-order nonlinear effects on CMOS-compatible nanophotonic platforms.


Inequality in prime-age adult deaths in a high AIDS mortality setting: does the measure of economic status matter?
We analyze deaths among prime-aged men and women during a 13-year period in a high AIDS mortality setting and examine the distribution of deaths by the economic status of these individuals at baseline using the 1991-2004 Kagera Health and Development Survey (KHDS). We investigate whether the distribution of subsequent prime-age adult deaths as measured by concentration indices depends on the measure of living standards used. We compare the performance of three measures: (1) per capita expenditure; (2) a modern wealth asset index replicating the asset index included in the 2004 Tanzanian AIDS Indicator Survey data file; and (3) a traditional wealth asset index, which includes only measures of traditional wealth. We find no evidence that economic status is linked to prime-age adult deaths, for both men and women, regardless of the measure of economic status used. This finding suggests both that more generally the measure of economic status used does not appear to be crucial, and specifically that relationships using traditional measures of wealth do not seem to differ from those using conventional measures.


VisANT: data-integrating visual framework for biological networks and modules.
VisANT is a web-based software framework for visualizing and analyzing many types of networks of biological interactions and associations. Networks are a useful computational tool for representing many types of biological data, such as biomolecular interactions, cellular pathways and functional modules. Given user-defined sets of interactions or groupings between genes or proteins, VisANT provides: (i) a visual interface for combining and annotating network data, (ii) supporting function and annotation data for different genomes from the Gene Ontology and KEGG databases and (iii) the statistical and analytical tools needed for extracting topological properties of the user-defined networks. Users can customize, modify, save and share network views with other users, and import basic network data representations from their own data sources, and from standard exchange formats such as PSI-MI and BioPAX. The software framework we employ also supports the development of more sophisticated visualization and analysis functions through its open API for Java-based plug-ins. VisANT is distributed freely via the web at http://visant.bu.edu and can also be downloaded for individual use.


Hierarchical aggregation for information visualization: overview, techniques, and design guidelines.
We present a model for building, visualizing, and interacting with multiscale representations of information visualization techniques using hierarchical aggregation. The motivation for this work is to make visual representations more visually scalable and less cluttered. The model allows for augmenting existing techniques with multiscale functionality, as well as for designing new visualization and interaction techniques that conform to this new class of visual representations. We give some examples of how to use the model for standard information visualization techniques such as scatterplots, parallel coordinates, and node-link diagrams, and discuss existing techniques that are based on hierarchical aggregation. This yields a set of design guidelines for aggregated visualizations. We also present a basic vocabulary of interaction techniques suitable for navigating these multiscale visualizations.


Epileptic seizures, coma and EEG burst-suppression from suicidal bupropion intoxication.
Bupropion, an amphetamine-like dual mechanism drug, is approved and increasingly used for the treatment of major depression, and its use is associated with a dose-dependent risk of epileptic seizures. Suicide attempts are frequent in major depression and often an overdose of the drugs available is ingested. Therefore, it is important to be aware of the clinical course, including EEG and neurological symptoms, as well as treatment and prognosis of bupropion intoxication. We report on the clinical and EEG course of a women who ingested 27 g of bupropion in a suicide attempt. Myoclonic seizures were followed by generalized tonic-clonic seizures and coma associated with EEG burst-suppression and brief tonic seizures. Active carbon and neuro-intensive care treatment, including respiratory support, were given. Within three days, the patient returned to a stable clinical condition with a mildly encephalopathic EEG. In conclusion, bupropion intoxication requires acute intensive care treatment and usually has a good prognosis, however, misinterpretation of the clinical and EEG presentation may lead to errors in management.


The demographic and biomedical case for late-life interventions in aging.
The social and medical costs of the biological aging process are high and will rise rapidly in coming decades, creating an enormous challenge to societies worldwide. In recent decades, researchers have expanded their understanding of the underlying deleterious structural and physiological changes (aging damage) that underlie the progressive functional impairments, declining health, and rising mortality of aging humans and other organisms and have been able to intervene in the process in model organisms, even late in life. To preempt a global aging crisis, we advocate an ambitious global initiative to translate these findings into interventions for aging humans, using three complementary approaches to retard, arrest, and even reverse aging damage, extending and even restoring the period of youthful health and functionality of older people.


Independent Living Oldest Old and Their Primary Health Provider: A Mixed Method Examination of the Influence of Patient Personality Characteristics.
This convergent mixed methods study examined 35 healthy, independent living individuals' (above 85 years) perceptions of their relationship with their primary health provider (PHP) and health practices. The relationship between PHP relationship perceptions and locus of control (LOC), resilience, and self-efficacy was explored through surveys and interviews. The majority indicated they visited their PHP just for preventative care; the number of PHP visits per year was significantly lower than reported for individuals above 85 by the CDC, possible reasons for this finding are provided. A positive relationship between LOC, resiliency, and self-efficacy for the oldest old was found. Few participants indicated their PHP had discussed normal changes with aging. This study has deepened understanding of the complexity inherent to the healthy oldest olds' relationship with their PHP. The findings suggest this relationship relates to the PHP's personal characteristics, the older adult patients' personality, and the influence of the accompanying patient escort.


The "cap-binding slot" of an mRNA cap-binding protein: quantitative effects of aromatic side chain choice in the double-stacking sandwich with cap.
The N7-methylguanine portion of the mRNA cap structure interacts with cap-binding proteins via an unusual double-stacking arrangement in which the positively charged cap is sandwiched between two parallel-oriented aromatic protein side chains. Three-dimensional costructures of cap with two mRNA cap-binding proteins, namely, translational initiation factor eIF4E and VP39 (the vaccinia virus-encoded mRNA cap-specific 2'-O-methyltransferase), have heretofore been reported. Despite striking similarities between the two proteins in the double stack with the cap, the stack differs most notably in the species of stacked side chain donated by the protein. Whereas eIF4E employs two tryptophans, VP39 uses a tyrosine and a phenylalanine. Here, we have generated tryptophan substitutions in VP39. Tryptophan substitution was shown, crystallographically, not to disrupt the maintenance of a bona fide parallel stack. However, the single-tryptophan and double-tryptophan substitutions were associated with increased affinity for cap nucleoside by factors of 10 and 50, respectively. VP39 interacted more strongly with a true substrate (containing portions of RNA downstream of the cap in addition to the cap itself) than with isolated cap nucleoside, by several orders of magnitude. VP39 mutants with tryptophan substitution at position 180 exhibited apparent defects in substrate catalytic rate during the first turnover cycle, indicating the possibility of an exquisite sensitivity of the catalytic center to subtle changes in substrate position brought about by alterations in the cap-binding slot. The X-ray structure of VP39 with a genuine nucleobase analogue of N7-methylguanosine, namely, N7,9-dimethylguanine, indicated that the N7-methylguanosine rotational orientation within the stack is a property of the cap nucleobase itself.


Diphthamide biosynthesis requires an organic radical generated by an iron-sulphur enzyme.
Archaeal and eukaryotic translation elongation factor 2 contain a unique post-translationally modified histidine residue called diphthamide, which is the target of diphtheria toxin. The biosynthesis of diphthamide was proposed to involve three steps, with the first being the formation of a C-C bond between the histidine residue and the 3-amino-3-carboxypropyl group of S-adenosyl-l-methionine (SAM). However, further details of the biosynthesis remain unknown. Here we present structural and biochemical evidence showing that the first step of diphthamide biosynthesis in the archaeon Pyrococcus horikoshii uses a novel iron-sulphur-cluster enzyme, Dph2. Dph2 is a homodimer and each of its monomers can bind a [4Fe-4S] cluster. Biochemical data suggest that unlike the enzymes in the radical SAM superfamily, Dph2 does not form the canonical 5'-deoxyadenosyl radical. Instead, it breaks the C(gamma,Met)-S bond of SAM and generates a 3-amino-3-carboxypropyl radical. Our results suggest that P. horikoshii Dph2 represents a previously unknown, SAM-dependent, [4Fe-4S]-containing enzyme that catalyses unprecedented chemistry.


Variation of the rfb gene clusters in Salmonella enterica.
In order to explore the genetic variation of O antigens of Salmonella enterica, we surveyed 164 strains (132 serovars) belonging to 45 serogroups, using 25 mostly single-gene rfb DNA probes for colony hybridization. The results revealed that strains within a serogroup have very similar or identical rfb genes. At least three of the four rhamnose genes were detected in all 17 serogroups reported to contain rhamnose, and one or more were detected in three others. The likelihood of being detected decreased in the order rfbB, rfbC, rfbA, and rfbD, which is the map order, suggesting a gradient of divergence. Mannose pathway genes were much less conserved, and of 27 groups reported to contain mannose or mannose derivatives colitose or fucose, only 9 hybridized to the rfbM and rfbK probes. Dideoxyhexose genes were found only in groups reported to contain dideoxyhexoses. Group D2, which had not been studied previously, appears to resemble group D1, with the substitution of one gene from group E1 to give a change in one linkage. In contrast to sugar pathway genes, sugar transferase genes did not in general hybridize to strains of other groups outside the closely related groups A, B, and D, with the exception of the galactose transferase gene also shared by groups C2, C3, and all E groups.


Plasma ceramide and glucosylceramide metabolism is altered in sporadic Parkinson's disease and associated with cognitive impairment: a pilot study.
Mutations in the gene coding for glucocerebrosidase (GBA), which metabolizes glucosylceramide (a monohexosylceramide) into glucose and ceramide, is the most common genetic risk factor for sporadic Parkinson's disease (PD). GBA mutation carriers are more likely to have an earlier age of onset and to develop cognitive impairment and dementia. We hypothesized that plasma levels of lipids involved in ceramide metabolism would also be altered in PD non-GBA mutation carriers and associated with worse cognition. Plasma ceramide, monohexosylceramide, and lactosylceramide levels in 26 cognitively normal PD patients, 26 PD patients with cognitive impairment or dementia, and 5 cognitively normal non-PD controls were determined by LC/ESI/MS/MS. Levels of all lipid species were higher in PD patients versus controls. Among PD patients, levels of ceramide C16:0, C18:0, C20:0, C22:0, and C24:1 and monohexosylceramide C16:0, C20:0 and C24:0 species were higher (all P<0.05) in those with versus without cognitive impairment. These results suggest that plasma ceramide and monohexosylceramide metabolism is altered in PD non-GBA mutation carriers and that higher levels are associated with worse cognition. Additional studies with larger sample sizes, including cognitively normal controls, are needed to confirm these findings.


The effect of saliva or serum on bacterial and Candida albicans colonization on type I collagen.
Colonization of Candida albicans on oral surfaces can serve as a reservoir for disseminated infections, such as aspiration pneumonia and gastrointestinal infection, particularly in the immunocompromised host. Therefore, the aim of this study was to investigate the effects of salivary and serum pellicles on C. albicans, Streptococcus mutans, S. sanguis, Lactobacillus and Actinomyces colonization on type I collagen, a major organic component of periodontal ligaments. The colonization potential of two isolates each of C. albicans, S. mutans and S. sanguis, and a single isolate each of Lactobacillus and Actinomyces to uncoated (control), saliva-coated or serum-coated type I collagen plates (surface area 143 mm(2), Cell Disk; Sumitomo, Tokyo, Japan) was examined using a bioluminescent adenosine triphosphate assay based on firefly luciferase-luciferin system. The results revealed that with mutans streptococci, a saliva pellicle was significantly more effective in promoting bacterial colonization compared with the pellicle-free collagen disc, and the serum-coated sample significantly inhibited the colonization of streptococci (anova; P < 0b01). In contrast, in the case of C. albicans, Lactobacillus and Actinomyces isolates, a serum pellicle was significantly more effective in promoting the colonization, followed by saliva pellicle and uncoated specimen (anova; P < 0b01). These results suggested that crevicular fluid rich in seruminous components would promote the colonization of Candida, Lactobacillus and Actinomyces on type I collagen as opposed to streptococci which showed greater avidity to saliva-coated collagen.


Decadal changes in habitat characteristics influence population trajectories of southern elephant seals.
Understanding divergent biological responses to climate change is important for predicting ecosystem level consequences. We use species habitat models to predict the winter foraging habitats of female southern elephant seals and investigate how changes in environmental variables within these habitats may be related to observed decreases in the Macquarie Island population. There were three main groups of seals that specialized in different ocean realms (the sub-Antarctic, the Ross Sea and the Victoria Land Coast). The physical and climate attributes (e.g. wind strength, sea surface height, ocean current strength) varied amongst the realms and also displayed different temporal trends over the last two to four decades. Most notably, sea ice extent increased on average in the Victoria Land realm while it decreased overall in the Ross Sea realm. Using a species distribution model relating mean residence times (time spent in each 50 × 50 km grid cell) to 9 climate and physical co-variates, we developed spatial predictions of residence time to identify the core regions used by the seals across the Southern Ocean from 120°E to 120°W. Population size at Macquarie Island was negatively correlated with ice concentration within the core habitat of seals using the Victoria Land Coast and the Ross Sea. Sea ice extent and concentration is predicted to continue to change in the Southern Ocean, having unknown consequences for the biota of the region. The proportion of Macquarie Island females (40%) utilizing the relatively stable sub-Antarctic region, may buffer this population against longer-term regional changes in habitat quality, but the Macquarie Island population has persistently decreased (-1.45% per annum) over seven decades indicating that environmental changes in the Antarctic are acting on the remaining 60% of the population to impose a long-term population decline in a top Southern Ocean predator.


Morphine for dyspnoea in chronic obstructive pulmonary disease: a before-after efficacy study.
Dyspnoea in patients with chronic obstructive pulmonary disease (COPD) is frequent and often persists despite conventional treatment. This study aimed to evaluate the efficacy and safety of oral morphine for dyspnoea in Japanese COPD patients. We conducted a multi-institutional, prospective, before-after study of morphine in COPD patients with dyspnoea at rest in seven hospitals. Patients received 12 mg of oral morphine per day (or 8 mg per day if low body weight or renal impairment). Primary outcome was change in the numerical rating scale (NRS) of current dyspnoea in the evening from Day 0 to Day 2. Secondary outcomes included changes in dyspnoea intensity in the evening from Day 0 to Day 1, dyspnoea intensity between the morning from Day 0 to Day 1 and Day 2, vital signs, nausea, somnolence, anorexia and other adverse events (AEs). A total of 35 patients were enrolled in this study between October 2014 and January 2018. One patient did not receive study treatment. Data from 34 patients was analysed. The NRS of dyspnoea intensity in the evening significantly decreased from 3.9 on Day 0 (95% CI: 3.1 to 4.8) to 2.4 on Day 2 (95% CI: 1.7 to 3.1; p=0.0002). Secondary outcomes significantly improved in a similar manner. There were no apparent changes in the mean scores of the opioid-related AEs and vital signs. One patient experienced grade 3 lung infection not associated with morphine. Other AEs were mild. Oral morphine is effective in alleviating dyspnoea in Japanese COPD patients. Trial registration UMIN000015288 (http://www.umin.ac.jp/ctr/index.htm).


PM2.5 constituents and hospital emergency-room visits in Shanghai, China.
Although ambient PM2.5 has been linked to adverse health effects, the chemical constituents that cause harm are largely unclear. Few prior studies in a developing country have reported the health impacts of PM2.5 constituents. In this study, we examined the short-term association between PM2.5 constituents and emergency room visits in Shanghai, China. We measured daily concentrations of PM2.5, organic carbon (OC), elemental carbon (EC), and eight water-soluble ions between January 1, 2011 and December 31, 2012. We analyzed the data using overdispersed generalized linear Poisson models. During our study period, the mean daily average concentration of PM2.5 in Shanghai was 55 μg/m(3). Major contributors to PM2.5 mass included OC, EC, sulfate, nitrate, and ammonium. For a 1-day lag, an interquartile range increment in PM2.5 mass (36.47 μg/m(3)) corresponded to 0.57% [95% confidence interval (CI): 0.13%, 1.01%] increase of emergency room visits. In all the three models used, we found significant positive associations of emergency room visits with OC and EC. Our findings suggest that PM2.5 constituents from the combustion of fossil fuel (e.g., OC and EC) may have an appreciable influence on the health impact attributable to PM2.5.


Neural responses in the macaque v1 to bar stimuli with various lengths presented on the blind spot.
Although there is no retinal input within the blind spot, it is filled with the same visual attributes as its surround. Earlier studies showed that neural responses are evoked at the retinotopic representation of the blind spot in the primary visual cortex (V1) when perceptual filling-in of a surface or completion of a bar occurs. To determine whether these neural responses correlate with perception, we recorded from V1 neurons whose receptive fields overlapped the blind spot. Bar stimuli of various lengths were presented at the blind spots of monkeys while they performed a fixation task. One end of the bar was fixed at a position outside the blind spot, and the position of the other end was varied. Perceived bar length was measured using a similar set of bar stimuli in human subjects. As long as one end of the bar was inside the blind spot, the perceived bar length remained constant, and when the bar exceeded the blind spot, perceptual completion occurred, and the perceived bar length increased substantially. Some V1 neurons of the monkey exhibited a significant increase in their activity when the bar exceeded the blind spot, even though the amount of the retinal stimulation increased only slightly. These response increases coincided with perceptual completion observed in human subjects and were much larger than would be expected from simple spatial summation and could not be explained by contextual modulation. We conclude that the completed bar appearing on the part of the receptive field embedded within the blind spot gave rise to the observed increase in neuronal activity.


Surprise as an explanation to auditory novelty distraction and post-error slowing.
Performance in sustained attention tasks is known to be slowed by the occurrence of unexpected task-irrelevant distractors (novelty distraction) and the detection of errors (posterror slowing), 2 well-established phenomena studied separately and regarded as reflecting distinct underpinning mechanisms. We measured novelty distraction and posterror slowing in an auditory-visual oddball task to test the hypothesis that they both involve an orienting response. Our results confirm that the 2 effects exhibit a positive interaction. We show that a trial-by-trial measure of surprise credibly accounts for our empirical data. We suggest that novelty distraction and posterror slowing both reflect an orienting response to unexpected events and a reappraisal of action plans. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Consumption and food utilization by individual larvae and the population of a wood borer Phymatodes maaki Kraatz (Coleoptera: Cerambycidae).
The consumption and the utilization of food by the individuals and the population of a wood borer, Phymatodes maaki Kraatz feeding on the dead wood of a Japanese wild vine, Vitis coignetiae Pulliat were studied at Mt. Daibosatsu, Yamanashi Pref., central Honshu from 1974 to 1976.As for individuals of P. maaki, the estimates of the ratio of growth to consumption and that of assimilation to consumption were very low, amounting to 1.24 and 3.23% in dry weight, 1.23 and 3.12% in carbon weight, and 16 and 13.6% in nitrogen weight respectively. The ratio of growth to assimilation was estimated at 35% in dry weight, 39% in carbon weight and 118% in nitrogen weight. Dry body weight of the adult was correlated with the amount of consumption of the larva.The parasitism by two Ichneumonidae species seemed to be the main mortality factor of the population of P. maaki. A minimum estimate of mortality from egg stage till emergence was 75%.As for the bioeconomy of the population, waste consumption i.e. the consumption of individuals that died before emergence amounted to 75% of total consumption. The growth efficiency (biomass of adults/total consumption) was 0.24% in dry weight, 0.26% in carbon weight and 3.4% in nitrogen weight. The waste efficiency (waste consumption/biomass of adults) was 310 in dry weight, 285 in carbon weight and 22 in nitrogen weight. Approximately 18% of the dead wood of V. coignetiae was decomposed by the feeding of the population.The dry weight of an adult was inversely correlated with density of larvae in the dead wood from which it emerged, but not significantly correlated with the diameter of the wood. The ratio of adult emergence i.e. the survival rate was inversely correlated with density of larvae, but the mortality by parasitism was not density dependent. This seems to suggest that there was an intraspecific competition for the limited food.


Estimates of hospitalization attributable to influenza and RSV in the US during 1997-2009, by age and risk status.
Estimates of influenza and respiratory syncytial virus (RSV) burden must be periodically updated to inform public health strategies. We estimated seasonal influenza- and RSV-attributable hospitalizations in the US from 1997 to 2009 according to age and risk status (NCT01599390). Multiple linear regression modelling was used to attribute hospitalizations to influenza or RSV using virological surveillance and hospitalization data. Hospitalization data were obtained from the US Nationwide Inpatient Sample and virology data were obtained from FluView (Centers for Disease Control and Prevention). Outcomes included any mention of ICD-coded respiratory disease and cardiorespiratory disease diagnoses. We also explored a broader definition of respiratory disease that included mention of relevant respiratory sign/symptoms and viral infection ("respiratory broad"). Applying the respiratory broad outcome, our model attributed ~300,000 and ~200,000 hospitalizations to influenza and RSV, respectively. Influenza A/H3N2 was the predominant cause of influenza-related hospitalizations in most seasons, except in three seasons when influenza B was dominant; likewise, A/H3N2 caused most influenza-related hospitalizations in all age segments, except in children <18 years where the relative contribution of A/H3N2 and B was similar. Most influenza A- and B-related hospitalizations occurred in seniors while approximately one half and one third of all RSV-related events occurred in children 0-4 years and seniors 65+ years, respectively. High-risk status was associated with higher risk of both influenza- and RSV-attributable hospitalizations in adults, but not in children. Our study assessed the burden of influenza and RSV, information that is important for both cost effectiveness studies and for prioritization of the development of antivirals and vaccines. For seniors, we found that the burdens of influenza and RSV were both substantial. Among children <18 years, about half of all influenza hospitalizations were due to influenza B, most occurring in children without noted risk conditions. RSV hospitalizations among children were confined to those 0-4 years. Our study also demonstrated the importance of the outcome used to estimate hospitalization burden. Our findings highlight the burden of influenza among children regardless of risk status and underscore the prevalence of RSV infections among both young children and older adults.


The MAL-ED cohort study: methods and lessons learned when assessing early child development and caregiving mediators in infants and young children in 8 low- and middle-income countries.
More epidemiological data are needed on risk and protective factors for child development. In The Etiology, Risk Factors and Interactions of Enteric Infections and Malnutrition and the Consequences for Child Health and Development (MAL-ED) cohort study, we assessed child development in a harmonious manner across 8 sites in Bangladesh, Brazil, India, Nepal, Pakistan, Peru, South Africa, and Tanzania. From birth to 24 months, development and language acquisition were assessed via the Bayley Scales of Infant and Toddler Development and a modified MacArthur Communicative Development Inventory. Other measures were infant temperament, the child's environment, maternal psychological adjustment, and maternal reasoning abilities. We developed standard operating procedures and used multiple techniques to ensure appropriate adaptation and quality assurance across the sites. Test adaptation required significant time and human resources but is essential for data quality; funders should support this step in future studies. At the end of this study, we will have a portfolio of culturally adapted instruments for child development studies with examination of psychometric properties of each tool used.


Notch1 mutations are drivers of oral tumorigenesis.
Disruption of NOTCH1 signaling was recently discovered in head and neck cancer. This study aims to evaluate NOTCH1 alterations in the progression of oral squamous cell carcinoma (OSCC) and compare the occurrence of these mutations in Chinese and Caucasian populations. We used a high-throughput PCR-based enrichment technology and next-generation sequencing (NGS) to sequence NOTCH1 in 144 samples collected in China. Forty-nine samples were normal oral mucosa from patients undergoing oral surgery, 45 were oral leukoplakia biopsies, and 50 were chemoradiation-naïve OSCC samples with 22 paired-normal tissues from the adjacent unaffected areas. NOTCH1 mutations were found in 54% of primary OSCC and 60% of premalignant lesions. Importantly, almost 60% of patients with leukoplakia with mutated NOTCH1 carried mutations that were also identified in OSCC, indicating an important role of these clonal events in the progression of early neoplasms. We then compared all known NOTCH1 mutations identified in Chinese patients with OSCC with those reported in Caucasians to date. Although we found obvious overlaps in critical regulatory NOTCH1 domains alterations and identified specific mutations shared by both groups, possible gain-of-function mutations were predominantly seen in Chinese population. Our findings demonstrate that premalignant lesions display NOTCH1 mutations at an early stage and are thus bona fide drivers of OSCC progression. Moreover, our results reveal that NOTCH1 promotes distinct tumorigenic mechanisms in patients from different ethnical populations.


Calibration of bioelectrical impedance analysis for body composition assessment in Ethiopian infants using air-displacement plethysmography.
Assessment of infant body composition (BC) is crucial to understand the consequences of suboptimal nutritional status and postnatal growth, and the effects of public health interventions. Bioelectrical impedance analysis (BIA) is a feasible, relatively inexpensive and noninvasive method for assessing BC. However, very little research has been conducted in low- and middle-income populations, where efforts to prevent or treat malnutrition in early life are a public health priority. We aimed to develop equations for predicting fat-free mass (FFM) and fat mass (FM) based on BIA in 0- to 6-month-old Ethiopian infants. The study comprised a total of 186 BC assessments performed in 101 healthy infants, delivered at Jimma University Specialized Hospital. Infant air-displacement plethysmography (IADP) was the criterion method, whereas weight, length, sex, age and an impedance index (L(2)/Z50) were predictors. Prediction equations were developed using stepwise multiple linear regression and the accuracy was evaluated with a 10-fold cross-validation approach. A linear regression model based on body weight, age and sex predicted FFM, estimated by IADP, with an adjusted R(2) and root mean square error (RMSE) of 0.94 and 200 g, respectively. Adding impedance index to the model resulted in a significantly improved model fit (R(2)=0.95; RMSE=181 g). For infants below 3 months of age, inclusion of impedance index did not contribute to an improved model fit for predicting FFM compared with a model already comprising weight, sex and age. The derived equations predicted FFM with acceptable accuracy and may be used in future field surveys, epidemiological studies and clinical trials conducted in similar sub-Saharan African population groups aged 0-6 months.


Red Blood Cell Distribution Width in Hospitalized COVID-19 Patients.
Introduction: Coronavirus disease-2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), is causing dramatic morbidity and mortality worldwide. The Red Blood Cell Distribution Width (RDW) has been strongly associated with increased morbidity and mortality in multiple diseases. Objective: To assess if elevated RDW is associated with unfavorable outcomes in hospitalized COVID-19. Methods: We retrospectively studied clinical outcomes of hospitalized COVID-19 patients for their RDW values. In-hospital mortality was defined as primary outcome, while septic shock, need for mechanical ventilation, and length of stay (LOS) were secondary outcomes. Results: A total of 294 COVID-19 patients were finally studied. Overall prevalence of increased RDW was 49.7% (146/294). RDW was associated with increased risk of in-hospital mortality (aOR, 4.6; 95%CI, 1.5-14.6) and septic shock (aOR, 4.6; 95%CI, 1.4-15.1) after adjusting for anemia, ferritin, lactate, and absolute lymphocyte count. The association remained unchanged even after adjusting for other clinical confounders such as age, sex, body mass index, coronary artery disease, hypertension, diabetes mellitus, and chronic obstructive pulmonary disease. No association was found instead with mechanical ventilation and median LOS. Conclusion: Elevated RDW in hospitalized COVID-19 patients is associated with a significantly increased risk of mortality and septic shock.


Structure and Function of the Acetylpolyamine Amidohydrolase from the Deep Earth Halophile Marinobacter subterrani.
Polyamines are small organic cations that are essential for cellular function in all kingdoms of life. Polyamine metabolism is regulated by enzyme-catalyzed acetylation-deacetylation cycles in a fashion similar to the epigenetic regulation of histone function in eukaryotes. Bacterial polyamine deacetylases are particularly intriguing, because these enzymes share the fold and function of eukaryotic histone deacetylases. Recently, acetylpolyamine amidohydrolase from the deep earth halophile Marinobacter subterrani (msAPAH) was described. This Zn2+-dependent deacetylase shares 53% amino acid sequence identity with the acetylpolyamine amidohydrolase from Mycoplana ramosa (mrAPAH) and 22% amino acid sequence identity with the catalytic domain of histone deacetylase 10 from Danio rerio (zebrafish; zHDAC10), the eukaryotic polyamine deacetylase. The X-ray crystal structure of msAPAH, determined in complexes with seven different inhibitors as well as the acetate coproduct, shows how the chemical strategy of Zn2+-dependent amide hydrolysis and the catalytic specificity for cationic polyamine substrates is conserved in a subterranean halophile. Structural comparisons with mrAPAH reveal that an array of aspartate and glutamate residues unique to msAPAH enable the binding of one or more Mg2+ ions in the active site and elsewhere on the protein surface. Notwithstanding these differences, activity assays with a panel of acetylpolyamine and acetyllysine substrates confirm that msAPAH is a broad-specificity polyamine deacetylase, much like mrAPAH. The broad substrate specificity contrasts with the narrow substrate specificity of zHDAC10, which is highly specific for N8-acetylspermidine hydrolysis. Notably, quaternary structural features govern the substrate specificity of msAPAH and mrAPAH, whereas tertiary structural features govern the substrate specificity of zHDAC10.


Relationship between renal dysfunction and outcomes in emergency department patients with potential acute coronary syndromes.
To determine whether patients with elevated creatinine who present to the emergency department (ED) with potential acute coronary syndrome (ACS) are at an increased risk of acute myocardial infarction (AMI) or 30-day cardiovascular (CV) events. A secondary analysis of a cohort study of patients presenting to the ED with potential ACS with serum creatinine measurements. Research assistants collected demographics, history, symptoms, hospital course and 30-day follow-up. Outcomes measured were in-hospital AMI and 30-day CV events (death, nonfatal AMI, revascularisation). Prespecified multivariable models included age, gender, race and cardiac risk factors and presenting electrocardiogram (ECG). We used a creatinine cut-off point of 132.6 mmol/l. Data are presented as OR and 95% CI. 3451 patients were enrolled (age, 52.9±13.2; 55% female patients; 64% black patients). There were 120 AMI during initial visit and 232 patients had 30-day CV events (43 deaths, 128 AMI, 120 revascularisations). Creatinine values were normal in 3086 (89.4%) and abnormal in 365 (10.5%) patients. In multivariable models the adjusted OR (95% CI) for the association between abnormal creatinine and AMI was 1.43 (0.88 to 2.30) and 30-day CV events was 1.57 (1.10 to 2.25). The odds of 30-day CV events were increased for patients who were older, male subjects, white, had hyperlipidaemia, hypertension or a history of CAD, or presented with an abnormal ECG. In patients with potential ACS in the ED, renal dysfunction predicts a higher likelihood of 30-day CV events, but not an independent predictor after controlling for other risk factors. It appears to be a marker of other CV risks.


Microbiome network analysis of co-occurrence patterns in anaerobic co-digestion of sewage sludge and food waste.
Addition of food waste (FW) as a co-substrate in anaerobic digesters of wastewater treatment plants is a desirable strategy towards achievement of the potential of wastewater treatment plants to become energy-neutral, diverting at the same time organic waste from landfills. Because substrate type is a driver of variations in phylogenetic structure of digester microbiomes, it is critical to understand how microbial communities respond to changes in substrate composition and concentration. In this work, high throughput sequencing was used to monitor the dynamics of microbiome changes in four parallel laboratory-scale anaerobic digesters treating sewage sludge during acclimation to an increasing amount of food waste. A co-occurrence network was constructed using data from 49 metagenomes sampled over the 161 days of the digesters' operation. More than half of the nodes in the network were clustered in two major modules, i.e. groups of highly interconnected taxa that had much fewer connections with taxa outside the group. The dynamics of co-occurrence networks evidenced shifts that occurred within microbial communities due to the addition of food waste in the co-digestion process. A diverse and reproducible group of hydrolytic and fermentative bacteria, syntrophic bacteria and methanogenic archaea appeared to grow in a concerted fashion to allow stable performance of anaerobic co-digestion at high FW.


The Role of Spirituality and Religiosity in Persons Living With Sickle Cell Disease: A Review of the Literature.
Sickle cell disease (SCD) is a serious debilitating chronic illness, affecting approximately 90,000 Americans and millions globally. Spirituality and religiosity (S/R) may ease the burden faced by persons living with SCD. The purpose of this study was to examine the role of S/R in adolescents and adults living with SCD in the research literature. The electronic databases Cumulative Index to Nursing and Allied Health Literature, Health Source Nursing/Academic, ProQuest Health Module, PsycINFO, Medline, PubMed, and the American Theological Library Association were searched from January 1995 to December 2014. Of the 89 studies retrieved, 11 articles between 2001 and 2013 met the inclusion criteria and were reviewed. Four themes emerged. The themes included (a) S/R as sources of coping, (b) S/R enhance pain management, (c) S/R influence health care utilization, and (d) S/R improve quality of life. Use of S/R may be significant in coping with SCD, managing pain, affecting hospitalizations, and affecting quality of life. This review can direct researchers exploring S/R in adolescents and adults living with SCD.


A stochastic, evolutionary model for range shifts and richness on tropical elevational gradients under Quaternary glacial cycles.
Quaternary glacial-interglacial cycles repeatedly forced thermal zones up and down the slopes of mountains, at all latitudes. Although no one doubts that these temperature cycles have left their signature on contemporary patterns of geography and phylogeny, the relative roles of ecology and evolution are not well understood, especially for the tropics. To explore key mechanisms and their interactions in the context of chance events, we constructed a geographical range-based, stochastic simulation model that incorporates speciation, anagenetic evolution, niche conservatism, range shifts and extinctions under late Quaternary temperature cycles along tropical elevational gradients. In the model, elevational patterns of species richness arise from the differential survival of founder lineages, consolidated by speciation and the inheritance of thermal niche characteristics. The model yields a surprisingly rich variety of realistic patterns of phylogeny and biogeography, including close matches to a variety of contemporary elevational richness profiles from an elevational transect in Costa Rica. Mountaintop extinctions during interglacials and lowland extinctions at glacial maxima favour mid-elevation lineages, especially under the constraints of niche conservatism. Asymmetry in temperature (greater duration of glacial than of interglacial episodes) and in lateral area (greater land area at low than at high elevations) have opposing effects on lowland extinctions and the elevational pattern of species richness in the model--and perhaps in nature, as well.


Dietary protein in weight management: a review proposing protein spread and change theories.
A large volume of human clinical data supports increased dietary protein for favorable changes to body composition, but not all data are conclusive. The aim of this review is to propose two theories, "protein spread theory" and "protein change theory" in an effort to explain discrepancies in the literature. Protein spread theory proposed that there must have been a sufficient spread or % difference in g/kg/day protein intake between groups during a protein intervention to see body composition and anthropometric differences. Protein change theory postulated that for the higher protein group, there must be a sufficient change from baseline g/kg/day protein intake to during study g/kg/day protein intake to see body composition and anthropometric benefits. Fifty-one studies met inclusion criteria. In studies where a higher protein intervention was deemed successful there was, on average, a 58.4% g/kg/day between group protein intake spread versus a 38.8% g/kg/day spread in studies where a higher protein diet was no more effective than control. The average change in habitual protein intake in studies showing higher protein to be more effective than control was +28.6% compared to +4.9% when additional protein was no more effective than control. Providing a sufficient deviation from habitual intake appears to be an important factor in determining the success of additional protein in weight management interventions. A modest increase in dietary protein favorably effects body composition during weight management interventions.


Promoted neuronal differentiation after activation of alpha4/beta2 nicotinic acetylcholine receptors in undifferentiated neural progenitors.
Neural progenitor is a generic term used for undifferentiated cell populations of neural stem, neuronal progenitor and glial progenitor cells with abilities for proliferation and differentiation. We have shown functional expression of ionotropic N-methyl-D-aspartate (NMDA) and gamma-aminobutyrate type-A receptors endowed to positively and negatively regulate subsequent neuronal differentiation in undifferentiated neural progenitors, respectively. In this study, we attempted to evaluate the possible functional expression of nicotinic acetylcholine receptor (nAChR) by undifferentiated neural progenitors prepared from neocortex of embryonic rodent brains. Reverse transcription polymerase chain reaction analysis revealed mRNA expression of particular nAChR subunits in undifferentiated rat and mouse progenitors prepared before and after the culture with epidermal growth factor under floating conditions. Sustained exposure to nicotine significantly inhibited the formation of neurospheres composed of clustered proliferating cells and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide reduction activity at a concentration range of 1 µM to 1 mM without affecting cell survival. In these rodent progenitors previously exposed to nicotine, marked promotion was invariably seen for subsequent differentiation into cells immunoreactive for a neuronal marker protein following the culture of dispersed cells under adherent conditions. Both effects of nicotine were significantly prevented by the heteromeric α4β2 nAChR subtype antagonists dihydro-β-erythroidine and 4-(5-ethoxy-3-pyridinyl)-N-methyl-(3E)-3-buten-1-amine, but not by the homomeric α7 nAChR subtype antagonist methyllycaconitine, in murine progenitors. Sustained exposure to nicotine preferentially increased the expression of Math1 among different basic helix-loop-helix proneural genes examined. In undifferentiated progenitors from embryonic mice defective of NMDA receptor subunit-1, nicotine was still effective in significantly inhibiting the proliferation. Functional α4β2 nAChR subtype would be constitutively expressed to play a role in the mechanism underlying the determination of proliferation and subsequent differentiation fate into a neuronal lineage in association with preferential promotion of Math1 expression in undifferentiated neural progenitors of developing rodent neocortex independently of NMDA receptor activation.


Multi-ancestry genome-wide association study of cannabis use disorder yields insight into disease biology and public health implications.
As recreational use of cannabis is being decriminalized in many places and medical use widely sanctioned, there are growing concerns about increases in cannabis use disorder (CanUD), which is associated with numerous medical comorbidities. Here we performed a genome-wide association study of CanUD in the Million Veteran Program (MVP), followed by meta-analysis in 1,054,365 individuals (ncases = 64,314) from four broad ancestries designated by the reference panel used for assignment (European n = 886,025, African n = 123,208, admixed American n = 38,289 and East Asian n = 6,843). Population-specific methods were applied to calculate single nucleotide polymorphism-based heritability within each ancestry. Statistically significant single nucleotide polymorphism-based heritability for CanUD was observed in all but the smallest population (East Asian). We discovered genome-wide significant loci unique to each ancestry: 22 in European, 2 each in African and East Asian, and 1 in admixed American ancestries. A genetically informed causal relationship analysis indicated a possible effect of genetic liability for CanUD on lung cancer risk, suggesting potential unanticipated future medical and psychiatric public health consequences that require further study to disentangle from other known risk factors such as cigarette smoking.


Safety considerations of plant polysaccharides for food use: a case study on phenolic-rich softwood galactoglucomannan extract.
A growing population and concern over the sufficiency of natural resources for feeding this population have motivated researchers and industries to search for alternative and complementary sources of food ingredients and additives. Numerous plant species and parts of plants are explored as raw materials for food production. An interesting example is wood; to date, only a few wood-based additives or ingredients are authorized for food use. Wood hemicelluloses, such as softwood galactoglucomannans (GGM), constitute an abundant bioresource that shows a high potential functionality in edible materials. Spruce GGM acts as a multi-functional emulsion stabilizer, and it could be used in various processed food products, replacing less effective, conventional emulsifiers. Before new materials can be released into the food market, their safety must be evaluated, according to the Novel Food regulation. This review focuses on the safety aspects that must be considered before polysaccharide- and phenolic-rich plant extracts can be awarded the status of authorized food ingredients. In this review, GGM is presented as a case study and examples are given of plant-based polysaccharides that are already authorized for food purposes. The legislation regarding Novel Food ingredients in Europe is also briefly reviewed.


A systematic survey of expression and function of zebrafish frizzled genes.
Wnt signaling is crucial for the regulation of numerous processes in development. Consistent with this, the gene families for both the ligands (Wnts) and receptors (Frizzleds) are very large. Surprisingly, while we have a reasonable understanding of the Wnt ligands likely to mediate specific Wnt-dependent processes, the corresponding receptors usually remain to be elucidated. Taking advantage of the zebrafish model's excellent genomic and genetic properties, we undertook a comprehensive analysis of the expression patterns of frizzled (fzd) genes in zebrafish. To explore their functions, we focused on testing their requirement in several developmental events known to be regulated by Wnt signaling, convergent extension movements of gastrulation, neural crest induction, and melanocyte specification. We found fourteen distinct fzd genes in the zebrafish genome. Systematic analysis of their expression patterns between 1-somite and 30 hours post-fertilization revealed complex, dynamic and overlapping expression patterns. This analysis demonstrated that only fzd3a, fzd9b, and fzd10 are expressed in the dorsal neural tube at stages corresponding to the timing of melanocyte specification. Surprisingly, however, morpholino knockdown of these, alone or in combination, gave no indication of reduction of melanocytes, suggesting the important involvement of untested fzds or another type of Wnt receptor in this process. Likewise, we found only fzd7b and fzd10 expressed at the border of the neural plate at stages appropriate for neural crest induction. However, neural crest markers were not reduced by knockdown of these receptors. Instead, these morpholino knockdown studies showed that fzd7a and fzd7b work co-operatively to regulate convergent extension movement during gastrulation. Furthermore, we show that the two fzd7 genes function together with fzd10 to regulate epiboly movements and mesoderm differentiation.


A one-enzyme PCR-RFLP assay for identification of six medically important Candida species.
Early identification of Candida isolates to the species level is necessary for effective antifungal therapy, and can also facilitate control of hospital infections. Phenotype-based methods for identifying Candida species are often difficult and time-consuming. Molecular biological techniques provide a useful alternative approach. In the present study, the ITS1-5.8S-ITS2 regions of fungal rRNA genes were amplified with universal primers in 20 standard strains. Digestion of the PCR products with one restriction enzyme, MspI, allowed discrimination of medically important Candida species, including C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, and C. guilliermondii. Using this method, we successfully identified 137 clinical isolates of Candida. Among them, C. albicans was identified as the most common species, followed by C. parapsilosis, C. tropicalis, C. glabrata, C. krusei, and C. guilliermondii. This method is a simple, rapid, and cost-effective method for differentiation between species that is applicable in clinical laboratories.


Skeletal muscle imaging and inflammatory myopathies.
A variety of modalities exist for the imaging of skeletal muscle including radiography, ultrasound, computed tomography, and MRI. This article highlights the utility of these modalities in evaluating skeletal muscle diseases. Newer techniques such as T2 mapping, blood oxygenation level dependent imaging, diffusion tensor imaging, and magnetic resonance spectroscopy are also explored. Cross-sectional imaging including computed tomography and magnetic resonance is the current standard in skeletal muscle imaging. The advantages of these modalities include the ability to image in different planes and to evaluate the distribution of disease and disease burden. Newer magnetic resonance-based techniques also provide functional information in addition to anatomic information. Radiography and ultrasound have a more limited role and are mainly used to detect calcifications and evaluate the texture of skeletal muscle. Magnetic resonance is a useful modality for evaluating skeletal muscle and allows for the assessment of disease burden. It can elucidate potential targets for biopsy. Newer magnetic resonance techniques can acquire functional information in addition to anatomic information and hold tremendous potential for detecting, characterizing, and monitoring treatment for inflammatory myopathies.


Reconstructing meaning through occupation after the death of a family member: accommodation, assimilation, and continuing bonds.
Reactions to death have been studied extensively from psychological, behavioral, and physiological perspectives. Occupational adaptation to loss has received scant attention. Qualitative research was undertaken to identify and describe occupational responses in bereavement. The constant comparative approach was used to analyze and interpret the occupational responses. Adaptive strategies of occupational accommodation and assimilation were used after the death of a family member. Desire to sustain bonds with the deceased motivated specific occupational engagements. These occupational responses served to reconstruct meaning after the death of a family member. These findings contribute to understanding adaptation after death by adding an occupational perspective to previous theories. Occupational therapists' abilities to support clients after loss can be enhanced through appreciation of occupational accommodation and assimilation and the role of continuing occupational bonds after the death of a loved one.


Osteoblast activity at the dental implant-bone interface: transmission electron microscopic and high voltage electron microscopic observations.
The purpose of this report is to present transmission electron microscopic and high voltage transmission electron microscopic (HVEM) observations of a longitudinal investigation examining the activities of osteoblasts and associated tissues apposing titanium and alumina oxide ceramic endosteal dental implants. The HVEM permitted 3-dimensional stereologic observations. All observations were obtained from undecalcified interfacial tissues from this in vivo experimental dog model using commercially available implants placed into the mandible. Two similar implants were placed in both sides of the mandible, with implants in 12 of the 18 dogs supporting fixed bridges for either 6 or 12 months. From the study, we observed that a mineralized matrix exists in direct apposition to the implant. Since bone does not interface the entire length of the implant, other interfacial zones were found to exist which consisted of unmineralized tissues. In such zones, we observed that osteoblasts were routinely found directly at the implant interface to the mandibular bone. These interfacial tissues included unmineralized collagen fibers, proteinaceous material, a finely fibrillar matrix, and the osteoblasts. This study has reinforced the concept that the oral tissue-dental implant interface is a dynamic zone consisting of remodeling activities of the osseous cells and extracellular matrices.


Increasing the diagnostic accuracy of medial temporal lobe atrophy in Alzheimer's disease.
Medial temporal lobe (MTL) atrophy is considered to be one of the most important predictors of Alzheimer's disease (AD). This study investigates whether atrophy in parietal and prefrontal areas increases the predictive value of MTL atrophy in three groups of different cognitive status. Seventy-five older adults were classified as cognitively stable (n = 38) or cognitively declining (n = 37) after three years follow-up. At follow-up, the grey matter of the MTL, inferior prefrontal cortex (IPC), and inferior parietal lobule (IPL) was delineated on MRI scans. Six years later, a dementia assessment resulted in distinguishing and separating a third group (n = 9) who can be considered as preclinical AD cases at scan time. Ordinal logistic regressions analysis showed that the left and right MTL, as well as the right IPC and IPL accurately predicted group membership. Receiver Operating Curves showed that the MTL was best in distinguishing cognitively stable from cognitively declining individuals. The accuracy of the differentiation between preclinical AD and cognitively stable participants improved when MTL and IPL volumes were combined, while differentiating preclinical AD and cognitively declined participants was accomplished most accurately by the combined volume of all three areas. We conclude that depending on the current cognitive status of an individual, adding IPL or IPC atrophy improved the accuracy of predicting conversion to AD by up to 22%. Diagnosis of preclinical AD may lead to more false positive outcomes if only the MTL atrophy is considered.


Effects of cigarette smoking on reproduction.
BACKGROUND Cigarette smoking is associated with lower fecundity rates, adverse reproductive outcomes and a higher risk of IVF failures. Over the last few decades, prevalence of smoking among women of reproductive age has increased. This review focuses on current knowledge of the potential effects of smoke toxicants on all reproductive stages and the consequences of smoke exposure on reproductive functions. METHODS We conducted a systematic review of the scientific literature on the impact of cigarette smoking and smoke constituents on the different stages of reproductive function, including epidemiological, clinical and experimental studies. We attempted to create hypotheses and find explanations for the deleterious effects of cigarette smoke observed in experimental studies. RESULTS Cigarette smoke contains several thousand components (e.g. nicotine, polycyclic aromatic hydrocarbons and cadmium) with diverse effects. Each stage of reproductive function, folliculogenesis, steroidogenesis, embryo transport, endometrial receptivity, endometrial angiogenesis, uterine blood flow and uterine myometrium is a target for cigarette smoke components. The effects of cigarette smoke are dose-dependent and are influenced by the presence of other toxic substances and hormonal status. Individual sensitivity, dose, time and type of exposure also play a role in the impact of smoke constituents on human fertility. CONCLUSIONS All stages of reproductive functions are targets of cigarette smoke toxicants. Further studies are necessary to better understand the deleterious effects of cigarette smoke compounds on the reproductive system in order to improve health care, help to reduce cigarette smoking and provide a better knowledge of the molecular mechanisms involved in reproductive toxicology.


[Cartographic notes on the work in the Family Health Strategy: relationships between workers and the population].
The objective of this intervention study was to map instituted and instituting movements present in the work of the Family Health Strategy in the development of their care practices. The theoretical framework is based on institutional analysis, using the schizoanalytic approach. Group meetings were carried out with the staff to discuss how they provided collective care in continuing health education. The study subjects were professionals from the team and students who were engaged in academic activity in the service. The average attendance was twelve people per meeting, and there were a total of eight meetings from March to July 2010. Data were grouped into two immanent strata: the relationships of the team and the relationship with clients. The strata point to the intersection of education and legal institutions and the social and technical division of labor. Collective thinking in groups appeared to be effective in denaturalizing established processes and interrogating places, knowledge and practices.


Investigating evolutionary perspective of carcinogenesis with single-cell transcriptome analysis.
We developed phase-switch microfluidic devices for molecular profiling of a large number of single cells. Whole genome microarrays and RNA-sequencing are commonly used to determine the expression levels of genes in cell lysates (a physical mix of millions of cells) for inferring gene functions. However, cellular heterogeneity becomes an inherent noise in the measurement of gene expression. The unique molecular characteristics of individual cells, as well as the temporal and quantitative information of gene expression in cells, are lost when averaged among all cells in cell lysates. Our single-cell technology overcomes this limitation and enables us to obtain a large number of single-cell transcriptomes from a population of cells. A collection of single-cell molecular profiles allows us to study carcinogenesis from an evolutionary perspective by treating cancer as a diverse population of cells with abnormal molecular characteristics. Because a cancer cell population contains cells at various stages of development toward drug resistance, clustering similar single-cell molecular profiles could reveal how drug-resistant sub-clones evolve during cancer treatment. Here, we discuss how single-cell transcriptome analysis technology could enable the study of carcinogenesis from an evolutionary perspective and the development of drug-resistance in leukemia. The single-cell transcriptome analysis reported here could have a direct and significant impact on current cancer treatments and future personalized cancer therapies.


Morphological variation on isolated populations of Praocis (Praocis) spinolai.
In this study, the morphological variations of four geographically isolated populations of Praocis (Praocis) spinolai Gay & Solier (Coleoptera: Tenebrionidae) in the transitional coastal desert, Chile, were studied. The study was conducted in the coastal area of Punta de Choros and Los Choros-Archipelago, which includes three islands: Choros, Damas, and Gaviota. 113 specimens of the species P. (P.) spinolai belonging to the four locations sampled were collected analyzed with geometric morphometrics techniques to explore the pattern of shape variation on the different isolated environments. The principal component analysis revealed a well-defined pattern of variation between the populations analyzed. Moreover, differences between populations emerged also from the canonical variation analysis and were confirmed by the Procrustes ANOVA. All analyses performed confirmed the existence of a pattern of variation, due to the isolation of the populations and to environmental effects. The islands are subject to more arid pressures than the continent, where there is a more stable environment and the presence of coastal wetlands and the coastal range of mountains act together and enable fog condensation. This study indicates the existence of a clear pattern of variation, which indicates an evolutionary trend among the population examined.


Blended vs. face-to-face cognitive behavioural treatment for major depression in specialized mental health care: study protocol of a randomized controlled cost-effectiveness trial.
Depression is a prevalent disorder, associated with a high disease burden and substantial societal, economic and personal costs. Cognitive behavioural treatment has been shown to provide adequate treatment for depression. By offering this treatment in a blended format, in which online and face-to-face treatment are combined, it might be possible to reduce the number of costly face-to-face sessions required to deliver the treatment protocol. This could improve the cost-effectiveness of treatment, while maintaining clinical effects. This protocol describes the design of a pilot study for the evaluation of the feasibility, acceptability and cost-effectiveness of blended cognitive behavioural therapy for patients with major depressive disorder in specialized outpatient mental health care. In a randomized controlled trial design, adult patients with major depressive disorder are allocated to either blended cognitive behavioural treatment or traditional face-to-face cognitive behavioural treatment (treatment as usual). We aim to recruit one hundred and fifty patients. Blended treatment will consist of ten face-to-face and nine online sessions provided alternately on a weekly basis. Traditional cognitive behavioural treatment will consist of twenty weekly sessions. Costs and effects are measured at baseline and after 10, 20 and 30 weeks. Evaluations are directed at cost-effectiveness (with depression severity and diagnostic status as outcomes), and cost-utility (with costs per quality adjusted life year, QALY, as outcome). Costs will encompass health care uptake costs and productivity losses due to absence from work and lower levels of efficiency while at work. Other measures of interest are mastery, working alliance, treatment preference at baseline, depressive cognitions, treatment satisfaction and system usability. The results of this pilot study will provide an initial insight into the feasibility and acceptability of blended cognitive behavioural treatment in terms of clinical and economic outcomes (proof of concept) in routine specialized mental health care settings, and an indication as to whether a well-powered clinical trial of blended cognitive behavioural treatment for depression in routine practice would be advisable. This will be determined based on the perspective of various stakeholders including patients, mental health service providers and health insurers. Strengths and limitations of the study are discussed. Netherlands Trial Register NTR4650 . Registered 18 June 2014.


A Comparative Study of MTA Solubility in Various Media.
Solubility of root filling materials is heavily influenced by the environment they are in contact with. This study compared the solubility of ProRoot MTA in deionized water and synthetic tissue fluid. Forty specimens of prepared MTA were immersed in deionized water and synthetic tissue fluid (20 samples each). The solubility was assessed after 7 and 28 days. Scanning electron microscope observation was also performed. The mean weight loss was evaluated using a digital scale. Data were analyzed using one-way ANOVA. Tukey test was performed for multiple comparisons. MTA solubility in synthetic tissue fluid was significantly lower than deionized water after 7 and 28 days (P<0.05). Secondary electron detectors revealed the presence of lumps and platelets on the surfaces of both specimens. Also, more voids were observed in specimen stored in deionized water. MTA dissolved faster in deionized water than synthetic tissue fluid. Despite this, the solubility of this material in both media was acceptable.


Estimating Individual Treatment Effect in Observational Data Using Random Forest Methods.
Estimation of individual treatment effect in observational data is complicated due to the challenges of confounding and selection bias. A useful inferential framework to address this is the counterfactual (potential outcomes) model, which takes the hypothetical stance of asking what if an individual had received both treatments. Making use of random forests (RF) within the counterfactual framework we estimate individual treatment effects by directly modeling the response. We find that accurate estimation of individual treatment effects is possible even in complex heterogenous settings but that the type of RF approach plays an important role in accuracy. Methods designed to be adaptive to confounding, when used in parallel with out-of-sample estimation, do best. One method found to be especially promising is counterfactual synthetic forests. We illustrate this new methodology by applying it to a large comparative effectiveness trial, Project Aware, to explore the role drug use plays in sexual risk. The analysis reveals important connections between risky behavior, drug usage, and sexual risk.


Comparison of physical activity between children with autism spectrum disorders and typically developing children.
Regular physical activity is important for promoting health and well-being; however, physical activity behaviors in children with autism spectrum disorders (ASD) have received little attention. We compared physical activity levels among 53 children with ASD and 58 typically developing children aged 3-11 years who participated in the Children's Activity and Meal Patterns Study (CHAMPS). After adjustment for age and sex the amount of time spent daily in moderate and vigorous activity was similar in children with ASD (50.0 minutes/day and typically developing children 57.1 minutes/day). However, parents reported that children with ASD participated in significantly fewer types of physical activities than did typically developing children (6.9 vs. 9.6, p <.0001) and spent less time annually participating in these activities than typically developing children (158 vs. 225 hours per year, p < 0.0001) after adjusting for age and sex. Although both groups of children engaged in similar levels of moderate and vigorous activity as measured by accelerometry, children with ASD engaged in fewer physical activities and for less time according to parental report, suggesting that some of the activity in children with ASD is not captured by standard questionnaire-based measures.


Relationship between ultrasound bone parameters, lung function, and body mass index in healthy student population.
Low bone mineral density has been reported in paediatric and adult patients with different lung diseases, but limited data are available on the association between lung function and bone density in a healthy young population. We explored the predictors of association between bone mass and pulmonary function in healthy first-year university students, focusing on body mass index (BMI). In this cross-sectional study we measured bone density with ultrasound and lung function with spirometry in 370 university students (271 girls and 99 boys). Information on lifestyle habits, such as physical activity, smoking, and alcohol consumption were obtained with a questionnaire. All lung function and bone parameters were significantly higher in boys than in girls (P<0.001). Underweight students had a significantly lower forced vital capacity (FVC%) (P=0.001 girls; P=0.012 boys), while overweight students had a significantly higher FVC% than normal weight students (P=0.024 girls; P=0.001 boys). BMI significantly correlated with FVC% (P=0.001) and forced expiratory volume in 1 second (FEV1 %) in both genders (P=0.001 girls; P=0.018 boys) and with broadband ultrasound attenuation (BUA) in boys. There were no significant associations between any of the bone and lung function parameters either in boys or girls. The most important determinant of lung function and ultrasound bone parameters in our study population was body mass index, with no direct association between bone density and lung function.


Mycoplasma capricolum subsp. capripneumoniae, the cause of contagious caprine pleuropneumonia, comprises two distinct biochemical groups.
Mycoplasma capricolum subsp. capripneumoniae, the cause of the World Organisation of Animal Health- listed contagious caprine pleuropneumonia, is a member of the Mycoplasma mycoides cluster which comprises five pathogenic mycoplasmas of ruminants. These mycoplasmas are closely related immunologically and genetically which can lead to difficulties for differential diagnosis. The patterns of substrate metabolism of strains of M. c. capripneumoniae, gathered from diverse geographic regions, were studied by measurement of oxygen uptake rates. The strains fell into two major biochemical groups: one which only oxidised organic acids and glycerol and the other which could additionally metabolise sugars. Furthermore when DNA-DNA hybridisation tests were carried out these two groups of strains could be separated by their degree of DNA homology, the mean hybridisation value between members of the two groups was 86% well above the value of 70% normally used to indicate separate species. DNA-DNA hybridisation was also carried out between M. c. capripneumoniae strains and other members of the M. mycoides cluster. These experiments used labelled DNA from two representative subsp. capripneumoniae strains; these were 7/1a (organic acid-oxidising) and 4/2 LC (glucose-oxidising). The results showed a particularly close relationship of the glucose-oxidising strain to M leachii strains.


Role of Lipoylation of the Immunodominant Epitope of Pyruvate Dehydrogenase Complex: Toward a Peptide-Based Diagnostic Assay for Primary Biliary Cirrhosis.
Primary biliary cirrhosis is an immune-mediated chronic liver disease whose diagnosis relies on the detection of serum antimitochondrial antibodies directed against a complex set of proteins, among which pyruvate dehydrogenase complex is considered the main autoantigen. We studied the immunological role of the lipoyl domain of this protein using synthetic lipoylated peptides, showing that the lipoyl chain chirality does not affect autoantibody recognition and, most importantly, confirming that both lipoylated and unlipoylated peptides are able to recognize specific autoantibodies in patients sera. In fact, 74% of patients sera recognize at least one of the tested peptides but very few positive sera recognized exclusively the lipoylated peptide, suggesting that the lipoamide moiety plays a marginal role within the autoreactive epitope. These results are supported by a conformational analysis showing that the lipoyl moiety of pyruvate dehydrogenase complex appears to be involved in hydrophobic interactions, which may limit its exposition and thus its contribution to the complex antigenic epitope. A preliminary analysis of the specificity of the two most active peptides indicates that they could be part of a panel of synthetic antigens collectively able to mimic in a simple immunoenzymatic assay the complex positivity pattern detected in immunofluorescence.


Molecular basis of proton block of L-type Ca2+ channels.
Hydrogen ions are important regulators of ion flux through voltage-gated Ca2+ channels but their site of action has been controversial. To identify molecular determinants of proton block of L-type Ca2+ channels, we combined site-directed mutagenesis and unitary current recordings from wild-type (WT) and mutant L-type Ca2+ channels expressed in Xenopus oocytes. WT channels in 150 mM K+ displayed two conductance states, deprotonated (140 pS) and protonated (45 pS), as found previously in native L-type Ca2+ channels. Proton block was altered in a unique fashion by mutation of each of the four P-region glutamates (EI-EIV) that form the locus of high affinity Ca2+ interaction. Glu(E)-->Gln(Q) substitution in either repeats I or III abolished the high-conductance state, as if the titration site had become permanently protonated. While the EIQ mutant displayed only an approximately 40 pS conductance, the EIIIQ mutant showed the approximately 40 pS conductance plus additional pH-sensitive transitions to an even lower conductance level. The EIVQ mutant exhibited the same deprotonated and protonated conductance states as WT, but with an accelerated rate of deprotonation. The EIIQ mutant was unusual in exhibiting three conductance states (approximately 145, 102, 50 pS, respectively). Occupancy of the low conductance state increased with external acidification, albeit much higher proton concentration was required than for WT. In contrast, the equilibrium between medium and high conductance levels was apparently pH-insensitive. We concluded that the protonation site in L-type Ca2+ channels lies within the pore and is formed by a combination of conserved P-region glutamates in repeats I, II, and III, acting in concert. EIV lies to the cytoplasmic side of the site but exerts an additional stabilizing influence on protonation, most likely via electrostatic interaction. These findings are likely to hold for all voltage-gated Ca2+ channels and provide a simple molecular explanation for the modulatory effect of H+ ions on open channel flux and the competition between H+ ions and permeant divalent cations. The characteristics of H+ interactions advanced our picture of the functional interplay between P-region glutamates, with important implications for the mechanism of Ca2+ selectivity and permeation.